Preparation method of daunorubicin hydrochloride rod-like crystal habit

Abstract

The invention discloses a preparation method of daunorubicin hydrochloride rod-like crystal habits, and belongs to the technical field of medicines. The method comprises the following steps: dissolving daunorubicin hydrochloride in a benign solvent, and heating and stirring until the daunorubicin hydrochloride is completely dissolved; and then adding a certain amount of poor solvent at a certain rate, then adding seed crystal, then adding a certain amount of poor solvent, controlling the stirring speed and cooling at a certain cooling rate after the addition is completed, continuing to maintain the stirring speed to grow the crystal, and filtering, washing and drying the product to obtain the daunorubicin hydrochloride rod-like crystal habit. The product crystal habit has the advantages of uniform particle size distribution, good product stability, good fluidity, large bulk density, high yield and the like. The method is high in experiment repeatability, is less interfered by external factors, and is suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and relates to a preparation method of a crystal habit of daunorubicin hydrochloride. BACKGROUND

[0002] Daunorubicin (also known as Zhen Dingmycin, Daunomycin, Rubidomycin, and Redmycin) belongs to an important antitumor drug in the anthracycline class of antibiotics, and has a chemical formula of C 27 H 29 NO 10 It is mainly used for acute lymphoblastic leukemia or acute myeloblastic leukemia that is resistant to commonly used antitumor drugs, but it is suitable to be used in combination with other drugs. Its main mechanism of action is in the process of nucleic acid synthesis of cells, can directly bind to DNA, hinder DNA synthesis and DNA-dependent RNA synthesis reaction, and has a particularly obvious effect on RNA, and can selectively act on purine nucleosides. The structural formula of daunorubicin is as follows:

[0003]

[0004] US6677309B1 and US6214330B1 patents protect the preparation method of daunorubicin. CN103641868B reports a method for producing daunorubicin hydrochloride from daunorubicin fermentation broth. The daunorubicin fermentation broth is first filtered by a porous metal membrane, the membrane filtration filtrate is cooled to 20-30℃, and then ultrafiltrated by a hollow fiber membrane assembly, the ultrafiltration filtrate is treated by a roll-type membrane nanofiltration, the obtained nanofiltration filtrate is cooled to 5-10℃, and then acetone is added for crystallization. After the crystals are completely precipitated, standing, solid-liquid separation is performed, and the obtained solid daunorubicin is obtained.

[0005] WO9929708A1 patent reports that daunorubicin is crystallized in a certain solvent system to obtain a plurality of crystal forms; one of the crystal forms has low moisture absorption, and the problem of solvent residue is solved.

[0006] CN115466295A patent provides a crystallization process for improving the crystal form of daunorubicin hydrochloride. The purity of daunorubicin hydrochloride before crystallization is improved by filtration, elution and other processes, and the adverse effects of impurities on crystallization are reduced. However, the process adopts 4 times of crystallization refining, which leads to the disadvantages of complex operation and high production cost.

[0007] Crystal habit refers to the macroscopic external shape of a crystal. The same substance can have different crystal habits under different crystallization conditions, and different substances can have the same crystal habit appearance. In the production process of crystal drugs, the crystal habit of drugs, as one of the important indicators of drugs, not only affects the post-processing process of the drugs such as filtration, washing and drying, but also affects the bulk density, fluidity and solubility of the drugs, thereby affecting the storage, transportation and use of the drugs. The properties of the drug itself and the physicochemical conditions of the crystal growth process are the two main factors that determine the morphology of the drug. Since the properties of the drug itself are difficult to change, the physicochemical environment of the crystal growth is usually changed by changing the crystallization conditions to obtain the target drug morphology. The present application is to better overcome the shortcomings of the prior art, suitable for industrial production, and improve the crystallization process of daunorubicin hydrochloride. SUMMARY

[0008] In order to overcome the shortcomings and deficiencies of daunorubicin hydrochloride in the prior art, the purpose of the present application is to provide a preparation method of rod-shaped crystal habit of daunorubicin hydrochloride. The method provided by the present application has the advantages of good stability, high bulk density, good fluidity, uniform particle size distribution and high yield.

[0009] In order to achieve the above-mentioned purpose, the present application realizes the following scheme:

[0010] Daunorubicin hydrochloride is dissolved in a good solvent, heated to T1, stirred to completely dissolve, a certain volume V1 of a poor solvent is added, a certain amount of crystal seeds is added, and then a certain volume V2 of the poor solvent is added at a rate v1. After the addition is completed, the stirring speed is controlled and a certain cooling rate is used to cool to T2. Continue to maintain the stirring speed and keep the crystal for a time t1. The crystal is filtered, washed and dried to obtain rod-shaped daunorubicin hydrochloride crystal.

[0011] In a preferred embodiment, the mass of daunorubicin hydrochloride to the volume of good solvent is 1:1-10 g / mL, preferably 1:2-5 g / mL.

[0012] In a preferred embodiment, the volume of the good solvent to the total volume (V1+V2) of the poor solvent is 1:4-10, preferably 1:6-7.

[0013] In a preferred embodiment, the good solvent is selected from one or a mixture of water and methanol; preferably methanol.

[0014] In a preferred embodiment, the poor solvent is selected from alcohols, esters, ethers, ketones, acetonitrile solvents other than methanol.

[0015] Among the alcohol, ethanol, isopropanol, n-butanol, ethylene glycol are preferred; among the ester, methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, methyl propionate, ethyl propionate, isopropyl propionate, butyl propionate are preferred; among the ether, tetrahydrofuran, 1,4-dioxane, diethyl ether are preferred; among the ketone, acetone, butanone, 2-pentanone, 3-pentanone, methyl vinyl ketone are preferred. Ethylene glycol and butanone are preferred. The good solvent and the poor solvent are preferably combined in a mutually soluble type.

[0016] In a preferred embodiment, the first addition of the poor solvent V1 is 5-40%, preferably 10-30% of the total amount of the poor solvent (V1+V2).

[0017] In a preferred embodiment, the rate of addition of the poor solvent v1 is 0.5-40 mL / min, preferably 5-25 mL / min.

[0018] In a preferred embodiment, the temperature T1 of the dissolving process is 30-90°C, preferably 40-80°C; and the temperature T2 of the temperature-lowering crystallization process is 0-30°C, preferably 10-25°C.

[0019] In a preferred embodiment, the amount of the seed crystal is 0.1-10.0 w.t.%, preferably 1.0-6.0 w.t.% of the amount of daunorubicin.

[0020] In a preferred embodiment, the stirring speed during the temperature-lowering to T2 and the seed crystal growing is controlled to be 50-600 rpm, preferably 100-300 rpm.

[0021] In a preferred embodiment, the stirring time t1 for the seed crystal growing is 1-12 h, preferably 2-8 h.

[0022] In a preferred embodiment, the temperature-lowering rate is 0.01-2°C / min, preferably 0.5-1.5°C / min.

[0023] In a preferred embodiment, the washing solvent is preferably a poor solvent with low boiling point, and the temperature of the washing solvent is controlled to be 0-20°C.

[0024] In a preferred embodiment, the drying temperature is 20-80°C, preferably 25-60°C.

[0025] In a preferred embodiment, the drying time is 2-24 h, preferably 4-12 h.

[0026] In terms of crystal habit, the purity of daunorubicin hydrochloride is required to be greater than 90%, preferably more than 95%.

[0027] In addition, the method of the present application can also be used for refining the crude daunorubicin hydrochloride.

[0028] The present invention has the following advantages and effects compared with the prior art:

[0029] (1) The preparation method of the present invention can be used to obtain rod-shaped crystal habit of daunorubicin hydrochloride. This crystal habit, i.e. appearance morphology, has the advantages of good production stability, high bulk density, good flowability, narrow particle size distribution, high purity and high yield.

[0030] (2) The experiment used PXRD (powder diffraction) as the basis for judging the crystal form of the product, and the spectra showed that the crystal forms were consistent. The experiment has strong repeatability and is less affected by external factors, making it particularly suitable for industrial production. Attached Figure Description

[0031] Figure 1 It is a needle-like crystal structure before the optimization of daunorubicin hydrochloride crystallization.

[0032] Figure 2 It is a rod-shaped crystal formed by the optimized crystallization of daunorubicin hydrochloride.

[0033] Figure 3 This is the PXRD pattern of daunorubicin hydrochloride before crystallization optimization.

[0034] Figure 4 This is the optimized PXRD pattern of daunorubicin hydrochloride crystals.

[0035] Figure 5 This is a particle size distribution diagram of daunorubicin hydrochloride crystals obtained in Comparative Example 1.

[0036] Figure 6 This is a particle size distribution diagram of the daunorubicin hydrochloride crystals obtained in Example 9. Detailed Implementation

[0037] The present invention will be further illustrated by the following embodiments. It should be understood that the embodiments of the present invention are merely for illustrating the present invention and are not intended to limit the present invention. Therefore, any simple improvements to the present invention under the premise of the method of the present invention are within the scope of protection claimed by the present invention.

[0038] In the following embodiments, various processes and methods not described in detail are conventional methods known in the art. The daunorubicin hydrochloride solid raw material used in Examples 1-9 of this invention has an HPLC purity of 99.30% and a crystal structure as described above. Figure 1 As shown. The crude daunorubicin hydrochloride used in Example 10 and Comparative Example 1 had an HPLC yield of 82.12%.

[0039] Example 1

[0040] Weigh 10g of daunorubicin hydrochloride solid, add 100mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 200mL of ethylene glycol (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 400mL of ethylene glycol (a poor solvent) dropwise at a rate of 5mL / min. After the addition is complete, control the stirring speed at 200rpm, cool to 25℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled ethanol (0-20℃), then dry at 50℃ for 10h to obtain 9.41g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 89.1% (HPLC: 99.79%).

[0041] Example 2

[0042] Weigh 10g of daunorubicin hydrochloride solid, add 50mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of ethylene glycol (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 300mL of ethylene glycol (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 200rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled ethanol (0-20℃), then dry at 50℃ for 10h to obtain 9.61g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 91.1% (HPLC: 99.77%).

[0043] Example 3

[0044] Weigh 10g of daunorubicin hydrochloride solid, add 20mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of ethylene glycol (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 150mL of ethylene glycol (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 200rpm, cool to 10℃ at a rate of 0.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled ethanol (0-20℃), then dry at 50℃ for 10h to obtain 9.11g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 86.1% (HPLC: 99.78%).

[0045] Example 4

[0046] Weigh 10g of daunorubicin hydrochloride solid, add 25mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 37.5mL of butanone (methyl ethyl ketone) solvent to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 112.5mL of butanone solvent dropwise at a rate of 15mL / min. After the addition is complete, control the stirring speed at 300rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 4h. Filter under reduced pressure and wash with a small amount of cooled butanone (0-20℃), then dry at 50℃ for 6h to obtain 9.31g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 88.1% (HPLC: 99.76%).

[0047] Example 5

[0048] Weigh 10g of daunorubicin hydrochloride solid, add 25mL of aqueous solvent, heat to 80℃, and stir until completely dissolved. Add 50mL of ethylene glycol (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 125mL of ethylene glycol (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 100rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled ethanol (0-20℃), then dry at 50℃ for 10h to obtain 9.68g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 91.8% (HPLC: 99.78%).

[0049] Example 6

[0050] Weigh 10g of daunorubicin hydrochloride solid, add 25mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of n-butanol (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 125mL of n-butanol (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 200rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled acetone (0-20℃), then dry at 50℃ for 8h to obtain 9.60g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 91.0% (HPLC: 99.79%).

[0051] Example 7

[0052] Weigh 10g of daunorubicin hydrochloride solid, add 25mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of acetone (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 350mL of acetone (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 100rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled acetone (0-20℃), then dry at 50℃ for 4h to obtain 9.29g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 87.9% (HPLC: 99.82%).

[0053] Example 8

[0054] Weigh 10g of daunorubicin hydrochloride solid, add 25mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of ethyl acetate (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 125mL of ethyl acetate (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 100rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled ethyl acetate (0-20℃), then dry at 50℃ for 3h to obtain 9.30g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 88.0% (HPLC: 99.83%).

[0055] Example 9

[0056] Weigh 10g of daunorubicin hydrochloride solid, add 25mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of tetrahydrofuran (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 125mL of tetrahydrofuran (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 100rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled tetrahydrofuran (0-20℃), then dry at 50℃ for 3h to obtain 9.26g of daunorubicin hydrochloride rod-shaped crystals, with a yield of 87.6% (HPLC: 99.85%).

[0057] Example 10

[0058] Weigh 10g of crude daunorubicin hydrochloride, add 25mL of methanol solvent, heat to 60℃, and stir until completely dissolved. Add 50mL of ethylene glycol (a poor solvent) to the system, then add 0.5g of daunorubicin hydrochloride seed crystals, stir for 1h, and then add 125mL of ethylene glycol (a poor solvent) dropwise at a rate of 10mL / min. After the addition is complete, control the stirring speed at 100rpm, cool to 15℃ at a rate of 1.5℃ / min, and maintain the stirring speed to grow crystals for 2h. Filter under reduced pressure and wash with a small amount of cooled ethanol (0-20℃), then dry at 50℃ for 10h to obtain 8.62g of rod-shaped crystals of daunorubicin hydrochloride, with a yield of 81.2% (HPLC: 99.72%).

[0059] Comparative Example 1

[0060] At room temperature, 10 g of crude daunorubicin hydrochloride was weighed and added to 125 mL of methanol solvent. The mixture was stirred until completely dissolved. Then, 1000 mL of n-butanol (a poor solvent) was added to the system, and the mixture was stirred to induce crystallization. The solution was filtered under reduced pressure and washed with a small amount of cooled acetone (0–20 °C). Finally, it was dried at 60 °C for 16 hours to obtain 8.21 g of needle-like crystals of daunorubicin hydrochloride (e.g., [missing information]). Figure 1 (acute needle-like crystals), yield 82.1%, HPLC: 97.23%.

[0061] The filtration time and drying time of the rod-shaped crystals and needle-shaped crystals of daunorubicin hydrochloride prepared in Examples 1-10 and the comparative examples were analyzed, as well as the solvent residue was analyzed according to the "Standard for daunorubicin hydrochloride, Part II, Chinese Pharmacopoeia 2020 Edition". The crystal parameters were also detected. The results are summarized in Tables 1 and 2.

[0062] Table 1:

[0063] Filtration time (min) Drying time (h) Solvent residue (GC) Example 1 6.5 10 Pass Example 2 6.5 10 Pass Example 3 6.5 10 Pass Example 4 5.0 6 Pass Example 5 6.5 10 Pass Example 6 6.0 8 Pass Example 7 3.0 4 Pass Example 8 3.5 3 Pass Example 9 4.0 3 Pass Example 10 6.5 10 Pass Comparative Example 1 30 16 Fail (n-butanol content greater than 1.0%)

[0064] Table 2

[0065]

[0066] Testing revealed that the rod-shaped crystals of daunorubicin hydrochloride obtained by this invention have a repose angle as low as 26 degrees, which is significantly better than the 45-degree repose angle of the needle-shaped crystals of daunorubicin hydrochloride obtained by the previous method. This indicates that the rod-shaped product of daunorubicin hydrochloride obtained by this invention has better flowability.

Claims

1. A method for preparing daunorubicin hydrochloride crystals, characterized in that, Daunorubicin hydrochloride was dissolved in a good solvent, heated to T1, and stirred until completely dissolved. A certain volume of poor solvent V1 was added, followed by a certain amount of seed crystals. Stirring continued, and then a certain volume of poor solvent V2 was added at a rate ν1. After the addition was complete, the stirring speed was controlled and the temperature was lowered to T2 at a certain cooling rate. The stirring speed was maintained for crystal growth time t1. The crystals were filtered, washed, and dried to obtain rod-shaped crystals of daunorubicin hydrochloride.

2. The preparation method according to claim 1, characterized in that, The mass ratio of daunorubicin hydrochloride to the volume ratio of the benign solution is 1:1 to 10 g / mL; preferably 1:2 to 5 g / mL.

3. The preparation method according to claim 1, characterized in that, The benign solvent is selected from one or a mixture of two solvents, namely water and methanol, with methanol being preferred.

4. The preparation method according to claim 1, characterized in that, The undesirable solvents are selected from alcohols, esters, ethers, ketones, and acetonitrile solvents other than methanol. Among them, alcohols are preferably ethanol, isopropanol, n-butanol, and ethylene glycol; esters are preferably methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, methyl propionate, ethyl propionate, isopropyl propionate, and butyl propionate; ethers are preferably tetrahydrofuran, 1,4-dioxane, and diethyl ether; and ketones are preferably acetone, butanone, 2-pentanone, 3-pentanone, and methyl vinyl ketone.

5. The preparation method according to claim 1, characterized in that, The undesirable solvents are selected from ethylene glycol and butanone.

6. The preparation method according to claim 1, characterized in that, The volume ratio of the good solvent to the total volume of the bad solvent, i.e., V1+V2, is 1:4 to 10; preferably 1:6 to 7; the V1 added in the first addition of the bad solvent accounts for 5 to 40% of the total volume of the bad solvent added, preferably 10 to 30%.

7. The preparation method according to claim 1, characterized in that, The rate of addition of the undesirable solvent ν1 is 0.5 mL / min to 40 mL / min, preferably 5 mL / min to 25 mL / min.

8. The preparation method according to claim 1, characterized in that, The temperature T1 is 30–90°C, preferably 40–80°C; the temperature T2 is 0–30°C, preferably 10–25°C.

9. The preparation method according to claim 1, characterized in that, The stirring speed is controlled at 50-600 rpm during cooling to T2 and during crystal growth; preferably 100-300 rpm.

10. The preparation method according to claim 1, characterized in that, The cooling rate is 0.01℃ / min to 2℃ / min, preferably 0.5℃ / min to 1.5℃ / min.

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